Reduced Representation Bisulfite Sequencing Library Preparation for Sperm DNA Methylation Analyses: Manual and Automated Methods.

Among epigenetic modifications, DNA methylation is extensively studied in sperm, and variations in sperm DNA methylation patterns have been associated with fertility, sperm quality, and in vitro fertilization outcomes in several species. Reduced representation bisulfite sequencing (RRBS) is a cost-effective method that has been successfully applied to sperm. However, RRBS library preparation remains sensitive and labor-intensive and can be subjected to diverse sources of technical variation. In order to improve RRBS library preparation performance, as well as its reproducibility, we adapted the reference protocol and implemented it on a Hamilton pipetting automaton.